Tissue-engineered three-dimensional model for tumor analysis

Abstract

A 3D decellularized bone scaffold seeded with cancer cells, such as prostate cancer cells or Ewing's sarcoma is provided. The three-dimensional includes Ewing's sarcoma (ES) tumor cells; and an engineered human bone scaffold. The engineered human bone scaffold further includes osteoblasts that secrete substance of the human bone, and osteoclasts that absorb bone tissue during growth and healing. The engineered human bone scaffold includes the tissue engineered three-dimensional model which recapitulates the osteolytic process. The engineered human bone scaffold is engineered by co-culturing of osteoblasts and osteoclasts. The osteoblast is produced by cell differentiation process from mesenchymal stem cells. The osteoclast is produced by cell differentiation from human monocytes, wherein the human monocytes are isolated from buffy coats. The scaffold can be used with cancer cell lines to identify therapeutic targets to slow, stop, and reverse tumor growth and progression as well as to predict the efficacy of potential therapeutics.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of International Application No. PCT / US2016 / 068478, filed Dec. 23, 2016, which claims priority to U.S. Provisional Application No. 62 / 387,121 filed Dec. 23, 2015, and a continuation-in-part of U.S. application Ser. No. 14 / 908,870, which is a continuation of International Application No. PCT / US14 / 49416, filed Aug. 1, 2014, which claims the benefit of U.S. Provisional Application No. 61 / 861,957, filed Aug. 2, 2013 and claims the benefit of U.S. Provisional Application No. 61 / 862,447, filed Aug. 5, 2013, all of which are incorporated by reference in their entirety herein.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]This invention was made with government support under grants EB002520 and EB17103 awarded by the NIH. The government has certain rights in the invention.BACKGROUNDTechnical Field[0003]The disclosed subject matter relates to providing a three-dimensional decellularized bone...

AI Technical Summary

Benefits of technology

[0021]In one embodiment, a cancer model is provided with a biomimetic microenvironment representing the pathophysiology of this malignancy. This is achieved by using three-dimensional (3D) instead of conventional two-dimensional (2D) cultures, with the aid of bone-engineering technology. In a 3D context, cancer cell lines modify their 2D transcriptional profile, recapitulating better the original tumor phenotype. This novel model is expected to be a powerful tool for predictive testing of anti-cancer and anti-metastatic compounds.

Problems solved by technology

The lack of ability to replicate in vitro the complex in vivo milieu of human cancer is a critical barrier to evaluation of the potential therapeutic targets for clinical application.

However, these 2D models fail to capture the true three-dimensional (3D) progression of tumors and are limited in their ability to identify therapeutic targets.

Our knowledge about the putative roles of the microenvironment on tumor exosomes is limited, due to a lack of experimental models that efficiently mimic the human in vivo situation.

Animal models used to study the effects of exosomes on cancer development often fail in representing the context of human disease.

The absence of physiological cell-cell and cell-matrix-interactions and the currently used non-physiological substrates cause disparity from the in vivo situation and lead to changes in cell morphology, proliferation and cellular processes, such as endo and exocytosis.

Although many drugs showed promise in these systems, most failed to translate into human patients and only ˜5% showed anti-tumor activity in clinical trials.

This discrepancy is caused by the lack of ability to sufficiently replicate the human microenvironment in these models.

Notably, ES cells are incapable of directly degrading bone matrix.

However, the crosstalk between tumor cells, osteoblasts and osteoclasts disrupts the bone remodeling and initiates either bone destruction (osteolytic tumors) or abnormal bone formation (osteoblastic tumors).

The lack of ability to replicate in vitro the bone osteolysis associated with the ES represents a critical barrier to understanding of the mechanisms underlying tumor progression and evaluating the new therapeutics.

However, modeling the bone invasion by cancer remains a challenge.

Most prostate cancer deaths are due to metastasis into bone, and yet there is not a good model of metastatic prostate cancer: in vitro, the cancer cells rapidly lose their cancer phenotype, and in vivo the mouse bone is not permissive for cancer cell invasion.

Unfortunately, the existing ES models have failed to faithfully predict human physiology and support the development of effective treatment modalities.

In spite of large investments, the use of these models has delayed drug discovery and exposed children to unnecessary chemicals, suggesting that modeling of the tumor progression requires interactions between tumor cells and their surrounding microenvironment.

However, these models are not able to mimic the disease.

Studies in animal models also have their limitations.

However, recapitulating tumor features in vitro is still a major challenge in the field.

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